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| M00770 | |
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| Product Description | Recombinant HEK293 cells stably express Angiotensin-converting enzyme 2 (ACE2) on the cell surface. The surface expression of ACE2 is validated by FACS analysis. This stable cell line product is designed for cell-based binding assays that measure the binding affinity and stability of antibody based biologics binding with ACE2, and can be utilized for SARS-Cov-2 pseudovirus neutralization assay for screening neutralizing antibodies. |
| Culture Properties | Adherent. |
| Stability | Stability: Stable through more than 20 passages with no significant changes in assay performance or expression profile. |
| Size | Two vials of frozen cells (>1×106 per vial in 1 mL). |
| Storage | Store cells in liquid nitrogen immediately upon receipt. Thaw and recover cells within one year from the date received. |
| Culture Medium | Culture Medium: DMEM, 10% FBS, 250 μg/ml Hygromycin B (Cat. No. 10687010, Life Technologies). |
| Complete Growth Medium | DMEM (Cat. No. 10569, Life Technologies), 10% FBS (Cat. No. 10099-141, Life Technologies) |
| Freeze Medium-DATA | 95% complete growth medium, 5% DMSO (Cat. No. D2650, Sigma). |
Figure 1. FACS analysis of cell surface expression of ACE2 on HEK293 cells. HEK293/ACE2 cells (red) and negative control HEK293 cells (green) were stained with SARS-CoV-2 Spike protein (RBD, His Tag) (Cat. No. Z03479, GenScript).
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Figure 2. Cell-based binding assay of SARS-CoV-2 spike protein RBD with HEK293/ACE2 cells. HEK293/ACE2 cells were incubated with SARS-CoV-2 Spike protein (RBD, His Tag) (Cat. No. Z03479, GenScript) in serial dilution. Then the cells were stained with THE™ His Tag Antibody [iFluor 488] Mouse (Cat. No. A01800, GenScript) and analyzed with flow cytometer. FACS analysis shows the EC50 of spike protein RBD binding with ACE2 on HEK293 cells is 1.07 ug/ml. »
Figure 3. Neutralization assay of SARS-CoV-2 spike protein RBD by ACE2-Fc fusion proteins on HEK293/ACE2 cells. HEK293/ACE2 cells were incubated with 1 µg/ml SARS-CoV-2 Spike protein (RBD, His Tag) (Cat. No. Z03479, GenScript) and ACE2-Fc fusion protein (Cat. No. Z03484, GenScript) in serial dilution. Then the cells were stained with THE™ His Tag Antibody [iFluor 488], (Cat. No. A01800, GenScript) and analyzed with flow cytometer. »
Figure 4. SARS-CoV-2 (B.1.1.529) pseudovirus infection on HEK293/ACE2 cells. SARS-CoV-2 pseudovirus is prepared with S glycoprotein as the envelope protein. The pseudovirus entry was initiated by recognition and binding between S glycoprotein and ACE2 on HEK293 cells.
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Figure 5. FACS analysis of cell surface expression of ACE2 on HEK293 cells of different passages. The HEK293/ACE2 cells and the negative control HEK293 cells were stained with Spike protein (RBD, His Tag) (Cat. No. Z03479, GenScript). P8, P13, P17 and P21 are cell passage numbers. »
For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.
